RESUMO
The study presents the killer functions of circulating neutrophils: myeloperoxidase activity, the ability to generate ROS, phagocytic activity, receptor status, NETosis, as well as the level of cytokines IL-2, IL-4, IL-6, IL-17A, and IL-18, granulocyte CSF, monocyte chemotactic protein 1, and neutrophil elastase in the serum of patients with uterine myoma and endometrial cancer (FIGO stages I-III). The phagocytic ability of neutrophils in uterine myoma was influenced by serum levels of granulocyte CSF and IL-2 in 54% of the total variance. The degranulation ability of neutrophils in endometrial cancer was determined by circulating IL-18 in 50% of the total variance. In uterine myoma, 66% of the total variance in neutrophil myeloperoxidase activity was explained by a model dependent on blood levels of IL-17A, IL-6, and IL-4. The risk of endometrial cancer increases when elevated levels of monocyte chemotactic protein 1 in circulating neutrophils are associated with reduced ability to capture particles via extracellular traps (96% probability).
Assuntos
Quimiocina CCL2 , Neoplasias do Endométrio , Interleucina-17 , Interleucina-6 , Neutrófilos , Humanos , Feminino , Neutrófilos/metabolismo , Neutrófilos/imunologia , Neoplasias do Endométrio/imunologia , Neoplasias do Endométrio/sangue , Neoplasias do Endométrio/patologia , Neoplasias do Endométrio/metabolismo , Interleucina-6/sangue , Quimiocina CCL2/sangue , Interleucina-17/sangue , Pessoa de Meia-Idade , Interleucina-4/sangue , Peroxidase/sangue , Peroxidase/metabolismo , Interleucina-18/sangue , Neoplasias Uterinas/sangue , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/patologia , Fator Estimulador de Colônias de Granulócitos/sangue , Fator Estimulador de Colônias de Granulócitos/metabolismo , Fagocitose , Leiomioma/sangue , Leiomioma/imunologia , Leiomioma/patologia , Leiomioma/metabolismo , Citocinas/sangue , Citocinas/metabolismo , Elastase de Leucócito/sangue , Elastase de Leucócito/metabolismo , Adulto , Armadilhas Extracelulares/metabolismo , Armadilhas Extracelulares/imunologia , Espécies Reativas de Oxigênio/metabolismo , Idoso , Interleucina-2RESUMO
OBJECTIVE: To examine the prevalence and prognostic role of tumor microenvironment (TME) markers in uterine carcinosarcoma (UCS) through immunohistochemical characterization. METHODS: The internal database of our institution was queried out for women with UCS who underwent surgery and thereafter postoperative chemotherapy with carboplatin and paclitaxel between January 2012 and December 2017. Tissue microarrays containing surgical samples of UCS from 57 women were assessed by immunohistochemistry for CD3, CD4, CD8, FOXP3, PD-1, PD-L1, and PD-L2. RESULTS: The mean age was 65.3 years (range, 49 to 79 years). For the epithelial component (E), CD3_E and CD4_E were highly expressed in 38 (66.7%) and in 40 (70.1%) patients, respectively, and were significantly associated with more advanced stages (p = 0.038 and p = 0.025, respectively). CD8_E was highly expressed in 42 (73.7%) patients, FOXP3_E 16 (28.1%), PD-1_E 35 (61.4%), PD-L1_E 27 (47.4%) and PD-L2_E 39 (68.4%). For the sarcomatous component (S), the prevalence of high expression was: CD3_S 6 (10.5%), CD4_S 20 (35.1%), CD8_S 44 (77.2%), FOXP3_S 8 (14%), PD-1_S 14 (24.6%), PD-L1_S 14 (24.6%) and PD-L2_S 8 (14%). By multivariate analysis, the CD8/FOXP3_S ratio (p = 0.026), CD4_E (p = 0.010), PD-L1_E (p = 0.013) and PD-L1_S (p = 0.008) markers significantly influenced progression-free survival. CD4/FOXP3_S ratio (p = 0.043), PD-1_E (p = 0.011), PD-L1_E (p = 0.036) and PD-L1_S (p = 0.028) had a significant association with overall survival. CONCLUSION: Some differences in UCS clinical outcomes may be due to the subtype of TILs and PD-1/PD-L1 axis immune checkpoint signaling.
Assuntos
Carcinossarcoma/imunologia , Carcinossarcoma/mortalidade , Linfócitos do Interstício Tumoral/metabolismo , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/mortalidade , Idoso , Antineoplásicos/uso terapêutico , Antígeno B7-H1/metabolismo , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/imunologia , Carboplatina/uso terapêutico , Carcinossarcoma/sangue , Feminino , Regulação Neoplásica da Expressão Gênica/imunologia , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Paclitaxel/uso terapêutico , Prevalência , Prognóstico , Receptor de Morte Celular Programada 1/metabolismo , Transdução de Sinais/imunologia , Microambiente Tumoral/imunologia , Neoplasias Uterinas/sangueRESUMO
OBJECTIVE: In 15% of patients with complete hydatidiform mole (CHM), disease progresses to post-molar gestational trophoblastic neoplasia (GTN) after curettage. Tumor infiltrating lymphocytes (TILs) are essential in overcoming disease in many tumors. Infiltrating lymphocyte composition and density may influence trophoblast regression and development of post-molar GTN. We analyzed immune cell composition and density in curettaged endometrium of patients with CHM which spontaneously regressed, and of patients with CHM which progressed to post-molar GTN. METHODS: Sixteen patients with CHM and spontaneous regression, and 16 patients with CHM which progressed to post-molar GTN were selected. Immune cell composition and density of natural killer (NK) cells, natural killer T (NKT)-like cells, Cytotoxic T cells, T-Regulatory and T-Helper cells, were determined by multiplex immunohistochemistry (mIHC). RESULTS: Curettaged endometrium of patients with CHM and spontaneous regression contained a slightly higher number of immune cells compared to patients with CHM which progressed to post-molar GTN. NKT-like cell density was significantly higher in patients with spontaneous regression compared to patients with CHM which progressed to post-molar GTN (483 ± 296 vs.295 ± 143 (mean ± SD), p = 0.03) respectively. NKT-like cell density in the spontaneous regression group was split in 'high' and 'low' (i.e. above and below the median number of NKT-like cells). In patients with high NKT-like cell density, hCG normalized earlier than in patients with low NKT-like cell density (9.5 weeks, (range 3.7-14) vs. 12.9 weeks, (range 8.6-17.9), p = 0.05). CONCLUSION: A high number of NKT-like cells in the endometrium of CHMs may contribute to spontaneous regression of molar trophoblast cells.
Assuntos
Endométrio/patologia , Mola Hidatiforme/imunologia , Linfócitos do Interstício Tumoral/imunologia , Células T Matadoras Naturais/imunologia , Neoplasias Uterinas/imunologia , Adulto , Gonadotropina Coriônica/sangue , Curetagem , Progressão da Doença , Endométrio/citologia , Endométrio/imunologia , Endométrio/cirurgia , Feminino , Citometria de Fluxo , Seguimentos , Idade Gestacional , Humanos , Mola Hidatiforme/sangue , Mola Hidatiforme/patologia , Mola Hidatiforme/cirurgia , Imunofenotipagem , Pessoa de Meia-Idade , Gravidez , Estudos Retrospectivos , Neoplasias Uterinas/sangue , Neoplasias Uterinas/patologia , Neoplasias Uterinas/cirurgia , Adulto JovemRESUMO
OBJECTIVE: Carcinosarcoma (CS) of the uterus or ovary is a rare, biphasic tumor comprising epithelial and mesenchymal elements, and exhibits more aggressive clinical features than its carcinoma counterpart. Four molecular subtypes of CS were recently established based on genomic aberration profiles (POLE, MSI, CNH, and CNL) and shown to be associated with multiple clinicopathological parameters, including patient outcomes. However, the role of the immune microenvironment in CS remains unclear. Here, we investigated the influence of the immune cells that infiltrate CS to better understand the immunological status of gynecological CS. METHODS: Tumor immune microenvironmental analyses on CS samples were performed using immune cell profiling with RNA-seq, transcriptomic subtyping with microenvironmental genes, and T-cell receptor repertoire assay. Carcinoma and sarcoma elements from CS samples were also assessed separately. RESULTS: Relying on estimations of tumor-infiltrating cell types from RNA-seq data, POLE and MSI (hypermutator) tumors showed an enrichment of M1 macrophages, plasma cells and CD8+ T cells, whereas CNH and CNL (non-hypermutator) tumors had high levels of M2 macrophages. Further subclassification by immune-related, non-cancer genes identified a fraction of tumors with distinct patient outcomes, particularly those with the CNH genomic aberration subtype. T-cell heterogeneity was independently correlated with prolonged progression-free survival. Differential analysis of carcinoma and sarcoma elements identified many shared mutations but there was little overlap in the T-cell receptor repertoire between the two elements. CONCLUSIONS: Tumor immune microenvironmental analyses could offer potential clinical utility in the stratification of gynecological CS above classification by genomic aberration subtype alone.
Assuntos
Carcinossarcoma/genética , Neoplasias Ovarianas/genética , Receptores de Antígenos de Linfócitos T/genética , Microambiente Tumoral/imunologia , Neoplasias Uterinas/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinossarcoma/imunologia , Carcinossarcoma/patologia , Estudos de Coortes , Biologia Computacional , Feminino , Heterogeneidade Genética , Humanos , Linfócitos do Interstício Tumoral , Pessoa de Meia-Idade , Mutação , Neoplasias Ovarianas/imunologia , Neoplasias Ovarianas/patologia , Ovário/imunologia , Ovário/patologia , Prognóstico , RNA-Seq , Microambiente Tumoral/genética , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/patologia , Útero/imunologia , Útero/patologia , Sequenciamento do ExomaRESUMO
PURPOSE: To evaluate radiotherapy-induced changes in the expression of programmed death ligand 1 (PD-L1), programmed death 1 (PD-1), and human leukocyte antigen class I (HLA-1) in patients with uterine cervical cancer, as well as infiltration of CD8+ and Forkhead box P3+ (FoxP3+) T lymphocytes into tumor tissue and the prognostic value of these parameters. MATERIALS AND METHODS: We performed immunohistochemical analysis of pre-radiotherapy biopsies and corresponding post-radiotherapy resected tissues in 104 uterine cervical cancer patients undergoing preoperative chemoradiotherapy or radiotherapy alone. We scored the expression of various proteins to distinguish positive from negative samples. RESULTS: PD-L1-expressing tumor cells (PD-L1 TC) increased significantly after chemoradiotherapy (pâ¯= 0.043). CD8+ T cell infiltration (pâ¯= 0.002) and FoxP3+ T cell infiltration (pâ¯= 0.003) decreased significantly after chemoradiotherapy. Expression of PD1, PD-L1-expressing immune cells (PD-L1 IC), and HLA1 did not change after chemoradiotherapy. In biopsy specimens obtained before chemoradiotherapy or radiotherapy, greater infiltration of CD8+ T cells (pâ¯= 0.001) and FoxP3+ T cells (pâ¯= 0.003) were significant predictors of better overall survival (OS). In surgical specimens obtained after chemoradiotherapy or radiotherapy, greater infiltration of PD-L1 TC was the only significant predictor of better OS (pâ¯< 0.001) and was related to a significantly lower probability of out-of-field recurrence (pâ¯= 0.005). CONCLUSION: Chemoradiotherapy induced an immunologic shift that increased PD-L1 TC. Chemoradiotherapy has immunological effects that can influence the results of treatment for uterine cervical cancer.
Assuntos
Antígeno B7-H1/análise , Carcinoma de Células Escamosas/radioterapia , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Antígenos HLA/análise , Terapia Neoadjuvante , Proteínas de Neoplasias/análise , Subpopulações de Linfócitos T/imunologia , Neoplasias Uterinas/radioterapia , Adulto , Idoso , Antineoplásicos Alquilantes/uso terapêutico , Linfócitos T CD8-Positivos/química , Linfócitos T CD8-Positivos/imunologia , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/cirurgia , Carcinoma de Células Escamosas/terapia , Quimiorradioterapia Adjuvante , Cisplatino/uso terapêutico , Feminino , Fatores de Transcrição Forkhead/análise , Humanos , Estimativa de Kaplan-Meier , Linfócitos do Interstício Tumoral/efeitos da radiação , Pessoa de Meia-Idade , Prognóstico , Modelos de Riscos Proporcionais , Curva ROC , Estudos Retrospectivos , Subpopulações de Linfócitos T/química , Resultado do Tratamento , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/cirurgia , Neoplasias Uterinas/terapiaRESUMO
OBJECTIVE: Uterine serous carcinoma (USC) is an aggressive variant of endometrial cancer with poor prognosis. Sacituzumab govitecan (SG) is a novel antibody-drug-conjugate (ADC) targeting trophoblast cell-surface antigen 2 (Trop-2), a transmembrane-calcium-signal-transducer, to deliver SN-38, the active metabolite of irinotecan. The objective of this study was to evaluate the expression of Trop-2 in USC and the preclinical activity of SG against primary USC cell-lines and xenografts. METHODS: We used immunohistochemistry (IHC) and flow-cytometry-based assays to evaluate Trop-2 expression and cell-viability in USC tissue and primary tumor-cell-lines after exposure to SG, non-targeting control ADC, and naked antibody hRS7-IgG. Antibody-dependent-cell-cytotoxicity (ADCC) against Trop-2+ and Trop-2- USC cell-lines was evaluated in vitro using 4-hr-Chromium-release-assays. In vivo activity of SG was tested against Trop-2+ USC xenografts by intravenous administration of SG, control ADC, and hRS7. RESULTS: Trop-2 expression by IHC was detected in 95.1% of USC samples (99/104). Primary tumor cell-lines overexpressing Trop-2 were significantly more sensitive to SG when compared to control ADC (p <0.05). Both SG and hRS7 mediated ADCC in Trop2+ USC cell-lines while no cytotoxicity was detected against Trop-2- cells. SG induced significant bystander killing of Trop-2- tumors when admixed with Trop-2+ tumors. SG caused growth-inhibition and increased survival in SG treated mice harboring Trop-2+ xenografts when compared to controls (p <0.05). CONCLUSIONS: SG is remarkably active against USC overexpressing Trop-2 in vitro and in vivo. Our results combined with SG clinical responses recently reported against multiple chemotherapy resistant human tumors further support clinical development of SG in USC patients with advanced/recurrent disease.
Assuntos
Anticorpos Monoclonais Humanizados/farmacologia , Antígenos de Neoplasias/imunologia , Camptotecina/análogos & derivados , Moléculas de Adesão Celular/imunologia , Imunoconjugados/farmacologia , Neoplasias Uterinas/tratamento farmacológico , Animais , Anticorpos Monoclonais Humanizados/imunologia , Citotoxicidade Celular Dependente de Anticorpos , Antígenos de Neoplasias/biossíntese , Camptotecina/imunologia , Camptotecina/farmacologia , Moléculas de Adesão Celular/biossíntese , Linhagem Celular Tumoral , Cistadenocarcinoma Seroso , Feminino , Citometria de Fluxo , Humanos , Imunoconjugados/imunologia , Imuno-Histoquímica , Camundongos , Camundongos SCID , Terapia de Alvo Molecular , Distribuição Aleatória , Análise Serial de Tecidos , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
An understanding of the interactions between immune cells and trophoblast cells, as well as choriocarcinoma cells, are of extreme importance in reproductive immunology and cancer immunology. In this study, we found that the human HLA-G-positive choriocarcinoma cell line JEG-3 upregulates CD4+CD25hiCD127lo T cells, increases the expression of HLA-G+CD4+ and CD8+ T cells, and decreases the expression of ILT2+ on CD4+ T cells in resting PBMCs after six days of co-culture. Expression of HLA-G on JEG-3 cells did not affect regulatory T cell phenotypes, but promoted modulation of pro-inflammatory cytokines IFN-γ, TNF-α and IL-17A. When JEG-3 cells were stimulated with rhIFN-γ prior to co-culture, CD4+HLA-G+ T cells were significantly increased, and IFN-γ and TNF-α elevated. Taken together, the results indicate that JEG-3 cells upregulate regulatory T cell phenotypes and modulate the level of pro-inflammatory cytokines, which might be important mechanisms in the tumor microenvironment and at the feto-maternal interface during pregnancy.
Assuntos
Coriocarcinoma/imunologia , Coriocarcinoma/metabolismo , Antígenos HLA-G/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linhagem Celular Tumoral/metabolismo , Citocinas/imunologia , Citocinas/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica/imunologia , Antígenos HLA-G/imunologia , Humanos , Interferon gama/metabolismo , Interleucina-17/metabolismo , Leucócitos Mononucleares/imunologia , Fenótipo , Gravidez , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Ativação Transcricional , Trofoblastos/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/metabolismoRESUMO
An aberrant p53 immunophenotype may be identified in several histotypes of endometrial carcinoma, and is accordingly recognized to lack diagnostic specificity in and of itself. However, based on the high frequency with which p53 aberrations have historically been identified in endometrial serous carcinoma, a mutation-type immunophenotype is considered to be highly sensitive for the histotype. Using an illustrative case study and a review of the literature, we explore a relatively routine diagnostic question: whether the negative predictive value of a wild-type p53 immunophenotype for serous carcinoma is absolute, that is, whether a p53-wild type immunophenotype is absolutely incompatible with a diagnosis of serous carcinoma. The case is an advanced stage endometrial carcinoma that was reproducibly classified by pathologists from 3 institutions as serous carcinoma based on its morphologic features. By immunohistochemistry, the tumor was p53-wild type (DO-7 clone), diffusely positive for p16 (block positivity), and showed retained expression of PTEN, MSH2, MSH6, MLH1, and PMS2. Next generation sequencing showed that there indeed was an underlying mutation in TP53 (D393fs*78, R213*). The tumor was microsatellite stable, had a low mutational burden (4 mutations per MB), and displayed no mutations in the exonuclease domain of DNA polymerase epsilon (POLE) gene. Other genomic alterations included RB1 mutation (R46fs*19), amplifications in MYST3 and CRKL, and ARID1A deletion (splice site 5125-94_5138del108). A review of the recent literature identified 5 studies in which a total of 259 cases of serous carcinoma were whole-exome sequenced. The average TP53 mutational rate in endometrial serous carcinoma was only 75% (range, 60 to 88). A total of 12 (33%) of 36 immunohistochemical studies reported a p53-aberrant rate of <80% in endometrial serous carcinoma. We discuss in detail several potential explanations that may underlie the scenario of serous carcinoma-like morphology combined with p53-wild-type immunophenotype, including analytic limitations, a nonserous histotype displaying morphologic mimicry of serous carcinoma, and true biological phenomena (including the possibility of a TP53-independent pathway of endometrial serous carcinogenesis). Ultimately, our central thematic question is provisionally answered in the negative. At present, the available data would not support a categorical conclusion that a p53 alteration is a necessary and obligate component in the genesis and/or diagnosis of endometrial serous carcinoma. On the basis of their collective experience, the authors proffer some recommendations on the use of p53 immunohistochemistry in the histotyping of endometrial carcinomas.
Assuntos
Biomarcadores Tumorais/genética , Neoplasias do Endométrio/patologia , Imunofenotipagem , Proteína Supressora de Tumor p53/imunologia , Neoplasias Uterinas/patologia , Feminino , Humanos , Pessoa de Meia-Idade , Proteína Supressora de Tumor p53/genética , Neoplasias Uterinas/imunologiaRESUMO
A 36-year-old with metastatic and refractory choriocarcinoma following single- and multi-agent chemotherapy and surgical metastectomy experienced a durable remission after receiving therapy with an anti-endoglin monoclonal antibody and bevacizumab. Treatment options and scientific advances in this disease are highlighted.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Coriocarcinoma/tratamento farmacológico , Neoplasias Uterinas/tratamento farmacológico , Adulto , Anticorpos Monoclonais/administração & dosagem , Bevacizumab/administração & dosagem , Coriocarcinoma/imunologia , Coriocarcinoma/patologia , Coriocarcinoma/cirurgia , Endoglina/antagonistas & inibidores , Endoglina/imunologia , Feminino , Doença Trofoblástica Gestacional/tratamento farmacológico , Doença Trofoblástica Gestacional/patologia , Doença Trofoblástica Gestacional/cirurgia , Humanos , Gravidez , Indução de Remissão , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/patologia , Neoplasias Uterinas/cirurgiaRESUMO
Therapeutic agents targeting the PD-1/PD-L1 axis have shown durable clinical responses in patients with various cancer types. Although objective responses are common, intrapatient heterogeneous responses have been described, and the mechanism for the different organ responses remains unknown. We present a series of patients in whom a lack of response was noted solely in the adrenal glands. This is the first case series describing 3 patients with heterogeneous patterns of response to pembrolizumab with progression of adrenal metastatic disease despite objective response (complete or partial response) in all other sites of metastatic disease. Two patients, one with melanoma and one with uterine carcinosarcoma, underwent robotic adrenalectomy for enlarging adrenal metastases. An additional patient with melanoma underwent laparotomy with attempted resection, but infiltration of the adrenal tumor into the inferior vena cava prohibited safe excision. This report provides additional insight into the heterogeneous patterns of disease response to anti-PD-1 therapy, highlighting the adrenal gland as a potential sanctuary site for this immunotherapy. These cases display the potential benefit of early surgical resection in this scenario and the pitfalls of delaying referral to a surgeon for assessment of operative intervention.
Assuntos
Neoplasias das Glândulas Suprarrenais/secundário , Glândulas Suprarrenais/patologia , Antineoplásicos Imunológicos/uso terapêutico , Carcinossarcoma/secundário , Melanoma/secundário , Neoplasias das Glândulas Suprarrenais/imunologia , Neoplasias das Glândulas Suprarrenais/cirurgia , Glândulas Suprarrenais/cirurgia , Adrenalectomia , Idoso , Anticorpos Monoclonais Humanizados/uso terapêutico , Carcinossarcoma/imunologia , Carcinossarcoma/terapia , Progressão da Doença , Feminino , Humanos , Melanoma/imunologia , Melanoma/terapia , Pessoa de Meia-Idade , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Receptor de Morte Celular Programada 1/imunologia , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologia , Resultado do Tratamento , Neoplasias Uterinas/tratamento farmacológico , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/patologiaRESUMO
We analyzed cytokine profile in blood serum of patients with uterine myoma and revealed significantly reduced level of IFNγ and a tendency towards a decrease in the levels of IL-1ß and TNFα; the levels of IL-6, IL-8, and IL-12p70 did not differ from those in healthy women. The drop in the concentrations of factors responsible for inflammation and angiogenesis in tissues are unfavorable for proliferation and differentiation of the uterine tissues.
Assuntos
Interferon gama/sangue , Interleucina-1beta/sangue , Leiomioma/sangue , Fator de Necrose Tumoral alfa/sangue , Neoplasias Uterinas/sangue , Útero/metabolismo , Adulto , Feminino , Expressão Gênica , Humanos , Inflamação , Interferon gama/genética , Interferon gama/imunologia , Interleucina-12/sangue , Interleucina-12/genética , Interleucina-12/imunologia , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-6/sangue , Interleucina-6/genética , Interleucina-6/imunologia , Interleucina-8/sangue , Interleucina-8/genética , Interleucina-8/imunologia , Leiomioma/imunologia , Leiomioma/patologia , Leiomioma/cirurgia , Pessoa de Meia-Idade , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Miomectomia Uterina , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/patologia , Neoplasias Uterinas/cirurgia , Útero/imunologia , Útero/patologia , Útero/cirurgiaRESUMO
OBJECTIVE: To investigate the growth mechanisms of large uterine leiomyoma (LULM) on the basis of a clinical morphology examination, by providing immunohistochemical (IHC) characteristics of the expression of growth factors (transforming growth factor-beta (TGFß) and platelet-derived endothelial cell growth factor (PD-ECGF)) and markers of stemness (CD117/c-kit, Connexin 43, Nestin) and proliferation (Ki-67). SUBJECT AND METHODS: The investigators examined surgical specimens from 38 women diagnosed with simple uterine leiomyoma (ULM), who had been divided into two groups: 1) 21 patients with LULM (>6 cm in diameter) (a study group); 2) 17 patients with small ULM (<4 cm in diameter) (a comparison group). Each group was also divided into two age subgroups (younger (<45 years) and older (≥45 years) subgroups (1a (n=12), 1b (n=9), 2a (n=8) and 2b (n=9), respectively. Histological specimens were used to make IHC examination with antibodies against TGFß, PD-ECGF, CD117/c-kit, Connexin 43, Nestin, and Ki-67. RESULTS: The growth mechanisms of LULM of simple histological structure were found to be associated with the larger number of growth zones in the tumors, with their enhanced cellular proliferative activity, and with the appearance of cells with signs of stemness, which is combined with the preserved subsequent maturation of tumor cells and determines the benign nature of LULM. CONCLUSION: There were differences in the molecular profile of LULM and small ULM, as well as LULM in perimenopausal and young women by the expression levels of Ki-67, TGFß, PD-ECGF, CD117, and Connexin 43, which can be used for diagnosis, prediction, and development of targeted therapies.
Assuntos
Leiomioma , Neoplasias Uterinas , Citocinas/análise , Feminino , Humanos , Imuno-Histoquímica , Leiomioma/imunologia , Leiomioma/patologia , Timidina Fosforilase , Fator de Crescimento Transformador beta , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/patologiaRESUMO
Uterine leiomyomas are benign tumors in the smooth muscle layer of the uterus. The most common histological type is the "usual leiomyoma", characterized by overexpression of ECM proteins, whereas the "cellular type" has higher cellular content. Our objective is to investigate the involvement of inflammatory and reparative processes in leiomyoma pathobiology. Using a morphological approach, we investigate the presence of inflammatory cells. Next, we determine the localization of the ECM, the presence/absence of fibrotic cells via α-sma and desmin and the immunohistochemical profile of the mesenchymal cells with respect to CD34. Finally, we explore the effect of inflammatory mediators (TNF-α, IL-1ß, IL-6, IL-15, GM-CSF and IFN-γ) on pro-fibrotic factor activin A mRNA expression in vitro. Higher numbers of macrophages were found inside and close to leiomyomas as compared to the more distant myometrium. Cellular leiomyomas showed more macrophages and mast cells than the "usual type". Inside the fibroid tissue, we found cells positive for α-sma, but negative for desmin and a large amount of collagen surrounding the nodule, suggestive of myofibroblasts producing ECM. In the myometrium and leiomyomas of the "usual type", we identified numerous CD34+ fibroblasts, which are known to give rise to myofibroblasts upon loss of CD34 expression. In leiomyomas of the "cellular type", stromal fibroblasts were CD34-negative. Finally, we found that TNF-α increased activin A mRNA in myometrial and leiomyoma cells. In conclusion, this study demonstrates the presence of inflammatory cells in uterine leiomyomas, which may contribute to excessive ECM production, tissue remodeling and leiomyoma growth.
Assuntos
Mediadores da Inflamação/metabolismo , Leiomioma Epitelioide/patologia , Miométrio/patologia , Neoplasias Uterinas/patologia , Útero/patologia , Actinas/metabolismo , Ativinas/imunologia , Antígenos CD34/metabolismo , Colágeno/metabolismo , Desmina/metabolismo , Feminino , Fibroblastos/metabolismo , Humanos , Imuno-Histoquímica , Inflamação/patologia , Leiomioma Epitelioide/imunologia , Macrófagos/imunologia , Mastócitos/imunologia , Miométrio/imunologia , RNA Mensageiro/metabolismo , Transdução de Sinais , Fator de Necrose Tumoral alfa/metabolismo , Neoplasias Uterinas/imunologiaRESUMO
BACKGROUND: Uterine serous carcinoma is an aggressive form of endometrial cancer that carries an extremely poor prognosis. Solitomab is a novel bispecific single-chain antibody construct that targets epithelial cell adhesion molecule on tumor cells and also contains a CD3 binding region. We evaluated the expression levels of epithelial cell adhesion molecule and the in vitro activity of solitomab against primary uterine serous carcinoma cell lines in vitro and ex-vivo in the ascites of patients with uterine serous carcinoma. OBJECTIVE: The purpose of this study was to determine the frequency of expression of epithelial cell adhesion molecule on uterine serous carcinoma cell lines and the ability of solitomab to modulate immune responses (T-cell proliferation, activation, cytokine production, and tumor killing) to tumor cells when it is combined with lymphocytes and epithelial cell adhesion molecule-positive cell lines or epithelial cell adhesion molecule-positive ascitic fluid in vitro. STUDY DESIGN: Epithelial cell adhesion molecule expression was evaluated by flow cytometry in a total of 14 primary uterine serous carcinoma cell lines. Sensitivity to solitomab-dependent cellular-cytotoxicity was tested against a panel of primary uterine serous carcinoma cell lines that express different levels of epithelial cell adhesion molecule in standard 4-hour chromium release assays. The proliferative activity, activation, cytokine secretion (ie, type I vs type II), and cytotoxicity of solitomab in autologous tumor-associated T cells in the ascitic fluid of patients with uterine serous carcinoma was also evaluated by carboxyfluorescein succinimidyl ester and flow-cytometry assays. Differences in epithelial cell adhesion molecule expression, solitomab-dependent cellular-cytotoxicity levels were analyzed with the use of an unpaired t test. T-cell activation marker increase and cytokine release were analyzed by a paired t test. RESULTS: Surface expression of epithelial cell adhesion molecule was found in 85.7% (12 of 14) of the uterine serous carcinoma cell lines that were tested by flow cytometry. Epithelial cell adhesion molecule-positive cell lines were found resistant to natural killer cells or T-cell-mediated killing after exposure to peripheral blood lymphocytes in 4-hour chromium-release assays (mean killing ± standard of the mean, 2.7% ± 3.1% after incubation of epithelial cell adhesion molecule-positive cell lines with control bispecific antibody construct). In contrast, after incubation with solitomab, epithelial cell adhesion molecule-positive uterine serous carcinoma cells became highly sensitive to T-cell cytotoxicity (mean killing, 25.7% ± 4.5%; P < .0001) by peripheral blood lymphocytes. Ex vivo incubation of autologous tumor-associated lymphocytes with epithelial cell adhesion molecule that expressed malignant cells in ascites with solitomab resulted in a significant increase in T-cell proliferation in both CD4+ and CD8+ T cells, increase in T-cell activation markers (ie, CD25 and HLA-DR), and a reduction in number of viable uterine serous carcinoma cells in ascites (P < .001). CONCLUSION: Solitomab induces robust immunologic responses in vitro that result in increased T-cell activation, proliferation, production of cytokines, and direct killing of tumor cells. These findings suggest that solitomab may represent a novel, potentially effective agent for the treatment of recurrent/metastatic and/or chemo-resistant uterine serous carcinoma-overexpressing epithelial cell adhesion molecule.
Assuntos
Anticorpos Biespecíficos/farmacologia , Antígenos de Neoplasias/efeitos dos fármacos , Antígenos de Neoplasias/imunologia , Antineoplásicos/farmacologia , Complexo CD3/imunologia , Carcinoma Papilar/tratamento farmacológico , Moléculas de Adesão Celular/efeitos dos fármacos , Moléculas de Adesão Celular/imunologia , Neoplasias Císticas, Mucinosas e Serosas/tratamento farmacológico , Neoplasias Uterinas/tratamento farmacológico , Antígenos de Neoplasias/análise , Líquido Ascítico/patologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Carcinoma Papilar/química , Carcinoma Papilar/imunologia , Moléculas de Adesão Celular/análise , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Citocinas/efeitos dos fármacos , Citocinas/metabolismo , Citotoxicidade Imunológica/efeitos dos fármacos , Molécula de Adesão da Célula Epitelial , Feminino , Citometria de Fluxo , Humanos , Ativação Linfocitária/efeitos dos fármacos , Neoplasias Císticas, Mucinosas e Serosas/química , Neoplasias Císticas, Mucinosas e Serosas/imunologia , Linfócitos T Citotóxicos/efeitos dos fármacos , Linfócitos T Citotóxicos/imunologia , Neoplasias Uterinas/química , Neoplasias Uterinas/imunologiaRESUMO
OBJECTIVE: To investigate the effects of the anti- ß2-glycoprotein I (anti-ß2-GPI) antibody on the tro- phoblast by evaluating the effects of the anti-ß2-GPI antibody on the expression of toll-like receptor (TLR) mRNA in choriocarcinoma cells and primary trophoblast cells. STUDY DESIGN: We cul- tured the choriocarcinoma cells (BeWo) and the pri- mary first trimester tropho- blast with the IgGs taken from anti-ß2-GPI antibody-positive and -negative sera. Four hours later we purified the RNAs from those cells. We measured the expressions of TLR mRNA in cells using real-time. PCR. RESULTS: The expression of TLR mRNA increased in BeWo cells and primary trophoblast cells cultured with the IgGs taken from anti-ß2-GPI antibody-positive women. Specifically, the expression of TLR1, 2, and 4 in BeWo cells -and TLRI, 3, 4, and 9 in first trimester trophoblast cells increased significantly. CONCLUSION: The anti-ß2-GPI antibody-positive IgG increased the TElR mRNA expression in choriocarcino- ma cells and primary trophoblast cells. We suggest that anti-ß2-GPI antibodies may bind to trophoblast and increase the expression of TLR mRNA.
Assuntos
Autoanticorpos/metabolismo , Coriocarcinoma/imunologia , RNA Mensageiro/metabolismo , Receptores Toll-Like/genética , Neoplasias Uterinas/imunologia , beta 2-Glicoproteína I/imunologia , Coriocarcinoma/metabolismo , Feminino , Humanos , Gravidez , Primeiro Trimestre da Gravidez , Receptores Toll-Like/metabolismo , Trofoblastos/imunologia , Trofoblastos/metabolismo , Células Tumorais Cultivadas , Neoplasias Uterinas/metabolismo , beta 2-Glicoproteína I/metabolismoRESUMO
BACKGROUND: While immunotherapy moved to the forefront of treatment of various cancers, it remains underexplored for uterine cancer. This might be due to the small patient population with advanced endometrial carcinoma and uterine sarcoma. Data about immunotherapeutic targets are scarce in endometrial carcinoma and lacking in uterine sarcoma. METHODS: Expression of five tumor-associated antigens (TAA) (BORIS, MUC1, hTERT, MAGE-A3 and Sp17) was validated in uterine tumor samples by immunohistochemistry (IHC) and/or quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). TAA immunogenicity was analyzed by determining spontaneous T cell responses towards overlapping peptide pools covering the whole TAA in patient blood. RESULTS: At mRNA level, MAGE-A3 and Sp17 were overexpressed in a minority of patients and BORIS was moderately overexpressed (26% in endometrial carcinoma and 62% in uterine sarcoma). hTERT was overexpressed in the vast majority of tumors. On protein level, MUC1 was upregulated in primary, recurrent and metastatic EMCAR and in metastatic US tumors. hTERT protein was highly expressed in both normal and malignant tissue. Spontaneous TAA-specific T cell responses were detected in a minority of patients, except for hTERT to which T cell responses occurred more frequently. CONCLUSIONS: These data point to MUC1 and hTERT as most suitable targets based on expression levels and T cell immunogenicity for use in immunotherapeutic regimens.
Assuntos
Adenocarcinoma/imunologia , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Sarcoma/imunologia , Linfócitos T/metabolismo , Neoplasias Uterinas/imunologia , Adenocarcinoma/genética , Antígenos de Superfície/genética , Antígenos de Superfície/metabolismo , Proteínas de Ligação a Calmodulina , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Linhagem Celular Tumoral , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imunoterapia , Técnicas In Vitro , Proteínas de Membrana , Mucina-1/genética , Mucina-1/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Peptídeos/farmacologia , Sarcoma/genética , Linfócitos T/efeitos dos fármacos , Telomerase/genética , Telomerase/metabolismo , Neoplasias Uterinas/genéticaRESUMO
Lymphangioleiomyomatosis (LAM) is a rare neoplastic metastatic disease affecting women of childbearing age. LAM is caused by hyperactivation of the mechanistic target of rapamycin complex 1 (mTORC1) as a consequence of tuberous sclerosis complex (TSC) 1/2 inactivation. Clinically, LAM results in cystic lung destruction. mTORC1 inhibition using rapamycin analogs (rapalogs) is partially effective in reducing disease progression and improving lung function. However, cessation of treatment results in continued progression of the disease. In the present study, we investigated the effectiveness of the combination of rapamycin treatment with resveratrol, an autophagy inhibitor, in the TSC2-null xenograft tumor model. We determined that this combination inhibits phosphatidylinositol-4,5-bisphosphate 3-kinase PI3K/Akt/mTORC1 signaling and activates apoptosis. Therefore, the combination of rapamycin and resveratrol may be an effective clinical strategy for treatment of LAM and other diseases with mTORC1 hyperactivation.
Assuntos
Antineoplásicos/farmacologia , Regulação Neoplásica da Expressão Gênica , Linfangioleiomiomatose/tratamento farmacológico , Sirolimo/farmacologia , Estilbenos/farmacologia , Proteínas Supressoras de Tumor/genética , Neoplasias Uterinas/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Quimioterapia Combinada , Feminino , Humanos , Linfangioleiomiomatose/genética , Linfangioleiomiomatose/imunologia , Linfangioleiomiomatose/patologia , Alvo Mecanístico do Complexo 1 de Rapamicina , Camundongos SCID , Complexos Multiproteicos/antagonistas & inibidores , Complexos Multiproteicos/genética , Complexos Multiproteicos/imunologia , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/imunologia , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/imunologia , Ratos , Resveratrol , Transdução de Sinais , Serina-Treonina Quinases TOR/antagonistas & inibidores , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/imunologia , Resultado do Tratamento , Proteína 1 do Complexo Esclerose Tuberosa , Proteína 2 do Complexo Esclerose Tuberosa , Proteínas Supressoras de Tumor/antagonistas & inibidores , Proteínas Supressoras de Tumor/deficiência , Proteínas Supressoras de Tumor/imunologia , Neoplasias Uterinas/genética , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/patologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The neonatal Fc receptor (FcRn) directs the transfer of maternal immunoglobulin G (IgG) antibodies across the placenta and thus provides the fetus and newborn with passive protective humoral immunity. Pathogenic maternal IgG antibodies will also be delivered via the placenta and can cause alloimmunity, which may be lethal. A novel strategy to control pathogenic antibodies would be administration of a nondestructive IgG antibody blocking antigen binding while retaining binding to FcRn. We report on 2 human IgG3 antibodies with a hinge deletion and a C131S point mutation (IgG3ΔHinge) that eliminate complement activation and binding to all classical Fcγ receptors (FcγRs) and to C1q while binding to FcRn is retained. Additionally, 1 of the antibodies has a single point mutation in the Fc (R435H) at the binding site for FcRn (IgG3ΔHinge:R435H). We compared transplacental transport with wild-type IgG1 and IgG3, and found transport across trophoblast-derived BeWo cells and ex vivo placenta perfusions with hierarchies as follows: IgG3ΔHinge:R435H>wild-type IgG1≥IgG3ΔHinge and IgG3ΔHinge:R435H=wild-type IgG1=wild-type IgG3>>>IgG3ΔHinge, respectively. Collectively, IgG3ΔHinge:R435H was transported efficiently from the maternal to the fetal placental compartment. Thus, IgG3ΔHinge:R435H may be a good candidate for transplacental delivery of a nondestructive antibody to the fetus to combat pathogenic antibodies.
Assuntos
Anticorpos/imunologia , Feto/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Imunoglobulina G/imunologia , Troca Materno-Fetal/imunologia , Placenta/imunologia , Receptores Fc/imunologia , Proteínas Recombinantes/imunologia , Anticorpos/metabolismo , Sítios de Ligação , Transporte Biológico , Coriocarcinoma/imunologia , Coriocarcinoma/metabolismo , Coriocarcinoma/patologia , Ensaio de Imunoadsorção Enzimática , Feminino , Feto/metabolismo , Citometria de Fluxo , Antígenos de Histocompatibilidade Classe I/metabolismo , Humanos , Imunoglobulina G/metabolismo , Recém-Nascido , Placenta/metabolismo , Gravidez , Receptores Fc/metabolismo , Proteínas Recombinantes/metabolismo , Neoplasias Uterinas/imunologia , Neoplasias Uterinas/metabolismo , Neoplasias Uterinas/patologiaRESUMO
Human papillomavirus (HPV) infection, particularly type 16, is causally associated with cancer of the uterine cervix, which mainly develops at the squamocolumnar (SC) junction. The progression of cervical HPV infections into (pre)neoplastic lesions suggests that viral antigens are not adequately recognized by innate immunity or presented to the adaptive immune system. Members of the defensin family have recently been found to inhibit viral and bacterial pathogens, to stimulate the migration of immune cells and to play a role in anticancer responses. In the present study, we focused on the poorly characterized human α-defensin 5 (HD-5) and its possible role in these processes. We showed that HD-5 was able to prevent HPV virion entry into cervical keratinocytes and to influence adaptive immunity. Indeed, this peptide specifically induced the chemoattraction and proliferation of both activated T lymphocytes and immature dendritic cells in a CCR2/CCR6-dependent manner and stimulated the infiltration of these professional antigen-presenting cells in a (pre)neoplastic epithelium transplanted in vivo in immunodeficient mice. No chemotactic effect was observed with plasmacytoid dendritic cells, macrophages or natural killer cells. Proliferative and angiogenic effects of HD-5 were also assessed in vitro and in vivo. However there was a striking regional disparity in expression of HD-5, being prominent in ectocervical, vaginal and vulvar neoplasia, while absent, or nearly so, in the cervical SC junction. Taken together, these results suggest one possible explanation for why the SC junction is uniquely vulnerable to both high-risk HPV infection (via reduced HD-5 expression and viral entry) and progression of neoplasia (via altered cell-mediated immune responses and altered microenvironment).
Assuntos
Colo do Útero/metabolismo , Infecções por Papillomavirus/imunologia , Lesões Pré-Cancerosas/imunologia , Neoplasias Uterinas/virologia , alfa-Defensinas/biossíntese , Animais , Western Blotting , Células Cultivadas , Colo do Útero/imunologia , Progressão da Doença , Ensaio de Imunoadsorção Enzimática , Feminino , Xenoenxertos , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Lesões Pré-Cancerosas/metabolismo , Lesões Pré-Cancerosas/virologia , Reação em Cadeia da Polimerase em Tempo Real , Neoplasias Uterinas/imunologia , Displasia do Colo do Útero/imunologia , Displasia do Colo do Útero/virologiaRESUMO
Uterine NK cells (uNK) play a role in the regulation of placentation, but their functions in nonpregnant endometrium are not understood. We have previously reported suppression of endometrial bleeding and alteration of spiral artery morphology in women exposed to asoprisnil, a progesterone receptor modulator. We now compare global endometrial gene expression in asoprisnil-treated versus control women, and we demonstrate a statistically significant reduction of genes in the IL-15 pathway, known to play a key role in uNK development and function. Suppression of IL-15 by asoprisnil was also observed at mRNA level (p < 0.05), and immunostaining for NK cell marker CD56 revealed a striking reduction of uNK in asoprisnil-treated endometrium (p < 0.001). IL-15 levels in normal endometrium are progesterone-responsive. Progesterone receptor (PR) positive stromal cells transcribe both IL-15 and IL-15RA. Thus, the response of stromal cells to progesterone will be to increase IL-15 trans-presentation to uNK, supporting their expansion and differentiation. In asoprisnil-treated endometrium, there is a marked downregulation of stromal PR expression and virtual absence of uNK. These novel findings indicate that the IL-15 pathway provides a missing link in the complex interplay among endometrial stromal cells, uNK, and spiral arteries affecting physiologic and pathologic endometrial bleeding.